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6th World Congress on Biotechnology

New Delhi, India

Janine Aparecida Correia Duraes Gandra

Janine Aparecida Correia Duraes Gandra

Novo Nordisk, Brazil

Title: Molecular and microbiological methods for Escherichia coli K12 traceability in the fermentation process

Biography

Biography: Janine Aparecida Correia Duraes Gandra

Abstract

The production of recombinant proteins in the fermentation process in biotechnology industry can be performed using genetically modified organism derived from Escherichia coli K12. The main health and safety concern on the use of GMOs (Genetically Modified Organism) is related to the potential of the gene transfers to other organisms, toxicity, inducing resistance to antibiotics and compromise biodiversity. The accidental release of GMOs in the recent years has increased the concern with safety and the need for methods able to detect them. Thus, policies of monitoring for disposal to the environment of bacterial carrying plasmids or other genetically modified organisms are required. In our research we associate the methods of membrane filtration and polymerase chain reaction (PCR) as tools for detection and quantification of E. coli K12 in downstream stages and in the final product of a fermentation process. Through contamination and recovery experiments it was possible to determine the detection limit for cells in CFU/mL following the membrane filtration method (1 CFU/mL). PCR reactions performed using E. coli presumptive identification colonies have proved that it is possible to detect E. coli K12 marker genes. Efforts have been undertaken to standardize and validate molecular methodologies in our laboratory among which PCR and more recently real-time PCR.